载紫杉醇脂质微泡联合超声破泡技术体外抑制多发性骨髓瘤细胞的研究

The inhibitory effect of paclitaxel-loaded lipid microbubbles combined with ultrasound on multiple myeloma cells in vitro

目的 观察载紫杉醇（PTX）的脂质微泡（MBs）联合超声（US）辐照体外作用于人多发性骨髓瘤U266细胞的效应.方法 采用机械振荡法制备PTX-MBs,镜下观察形态,检测其粒径、载药率、包封率和稳定性.检测单纯超声对细胞增殖和凋亡的影响.设Control组（Ⅰ）、PTX组（Ⅱ）、PTX＋ US组（Ⅲ）、PTX-MBs组（Ⅳ）、PTX-MBs＋ US组（Ⅴ）,细胞计数试剂盒（CCK-8）法检测细胞的存活率,流式细胞术检测细胞的凋亡,软琼脂克隆实验检测细胞的克隆形成率.结果 PTX-MBs平均粒径为（353.9±25.7） nm,包封率为（86.25±7.31）％,载药率为（20.09±3.46）％.0.5 W/cm2的超声作用40 s对细胞增殖和凋亡无明显影响.各实验组中,Ⅴ组的细胞存活率[（32.89±5.59）％]较其他组[Ⅰ组：（98.42±1.47）％,Ⅱ组：（63.41±2.42）％,Ⅲ组：（58.79±4.16）％,Ⅳ组：（93.39±3.21）％]明显降低（P＜0.05）,Ⅴ组的细胞凋亡率[（28.48±3.89）％]较其他组[Ⅰ组：（3.03±1.09）％,Ⅱ组：（16.69±3.14）％,Ⅲ组：（16.84±1.11）％,Ⅳ组：（4.43±1.58）％]明显增强（P＜0.05）,Ⅴ组细胞的软琼脂克隆形成率[（6.00±1.00）％]较其他组[Ⅰ组：（35.00±5.00）％,Ⅱ组：（23.67±1.53）％,Ⅲ组：（20.00±2.65）％,Ⅳ组：（32.33±5.03）％]明显降低（P＜0.05）.结论 PTX-MBs是良好的药物传递载体,联合超声辐照体外作用于多发性骨髓瘤细胞可显著抑制细胞增殖、诱导细胞凋亡以及抑制细胞克隆形成.

Objective To prepare paclitaxel-loaded lipid microbubbles （PTX-MBs） and investigate the effect of the MBs combined with ultrasound on multiple myeloma U266 cells in vitro.Methods PTX-MBs prepared by mechanic vibration method were observed under microscope.The size distribution, drug-loading efficiency and encapsulation efficiency were further detected.Ultrasound （US） was simply applied to multiple myeloma cells to observe the inhibitory effects.Cells were divided into Control group （Ⅰ）, PTX group（Ⅱ）, PTX ＋ US group（Ⅲ）, PTX-MBs group（Ⅳ）, PTX-MBs ＋ US group（Ⅴ）.The inhibitory effect on cell proliferation was detected by cell counting kit-8 （CCK-8） method, the cell apoptosis was detected by flow cytometry （FCM）, and the clone formation rate was detected by soft agar cloning method.Results PTX-MBs has good dispersion with the size [（353.9 ± 25.7） nm], encapsulation efficiency [（86.25 ± 7.31） %], and drug-loading efficiency [（20.09 ± 3.46） %].Ultrasound at 0.5 W/cm2 for40 s did no harm to cells.The survival rate was lower in PTX-MBs ＋ US group[（32.89 ± 5.59） %] than in other groups [Group Ⅰ ： （98.42 ± 1.47） %, Group Ⅱ ： （63.41 ± 2.42） %, Group Ⅲ ： （58.79 ±4.16）%, Group Ⅳ： （93.39 ±3.21）%, P〈0.05].The apoptosis rate was higher in PTX-MBs ＋ US group [（28.48 ± 3.89） %] than in other groups [Group Ⅰ ： （3.03 ± 1.09） %, Group Ⅱ ： （16.69 ± 3.14） %, Group Ⅲ： （16.84 ± 1.11） %, Group Ⅳ ： （4.43 ± 1.58） %, P 〈 0.05].The clone formation rate was higher in PTX-MBs ＋ US group [（6.00 ± 1.00） %] than in other groups [Group Ⅰ ： （35.00 ± 5.00） %, Group Ⅱ ： （23.67 ± 1.53） %, Group Ⅲ ： （20.00 ± 2.65） %, Group Ⅳ ： （32.33 ± 5.03） %, P 〈 0.05].Conclusion As drug delivery system, PTX-MBs can inhibit the proliferation, induce the apoptosis and inhibit the clone formation of multiple myeloma cells when combined with ultrasound.